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目的探讨赖氨大黄酸对HeLa细胞周期的影响及其机制。方法 MTT法检测赖氨大黄酸对HeLa细胞增殖的影响;流式细胞技术检测活性氧水平和细胞周期变化;显微镜下观察HeLa细胞形态变化;Western blot法检测p53和p21蛋白表达。结果赖氨大黄酸剂量依赖性地抑制HeLa细胞增殖,IC50约为83μmol·L-1。随着赖氨大黄酸剂量的增加,HeLa细胞中活性氧水平增加,并出现S期和G2期阻滞,以及细胞空泡样变性。通过Western blot研究发现赖氨大黄酸处理后的p53蛋白磷酸化水平呈剂量依赖性地增加,同时下游周期相关蛋白p21表达水平也随之增加,而p53蛋白表达水平未出现显著地剂量依赖性变化。结论赖氨大黄酸通过增加HeLa细胞中活性氧水平,引起细胞周期相关蛋白p53的磷酸化水平和p21蛋白表达水平增加,引起HeLa细胞出现空泡样变性,并诱导S期和G2期阻滞。
Objective To investigate the effect of lysine on HeLa cell cycle and its mechanism. Methods MTT assay was used to examine the effect of rhein on proliferation of HeLa cells. Flow cytometry (FCM) was used to detect ROS levels and cell cycle changes. Morphological changes of HeLa cells were observed under microscope. The expressions of p53 and p21 protein were detected by Western blot. Results Larch could inhibit the proliferation of HeLa cells in a dose-dependent manner with an IC50 of 83μmol·L-1. As the dose of lysine increased, the level of reactive oxygen species in HeLa cells increased, and S phase and G2 phase arrest, as well as vacuolar degeneration of cells. Western blot analysis showed that the phosphorylation of p53 protein increased in a dose-dependent manner, and the expression of downstream protein p21 also increased, while the expression of p53 protein did not show significant dose-dependent changes . Conclusion L-Rhein can induce the vacuolar degeneration of HeLa cells and induce the arrest of S phase and G2 phase by increasing the level of ROS in HeLa cells, resulting in the increase of phosphorylation of p53 and the expression of p21 protein.